This invention relates to novel bioactive peptide compositions which enhance the growth of animals, a process for the preparation of same and the use of such compositions for enhancing the growth of warmblooded animals and fish.
It is well known that porcine and bovine plasma protein can be used in feed formulations for warm blooded animals to improve health, growth and general performance. However, the need for a replacement for such plasma protein is of great importance. The cost of plasma protein continues to increase as foreign and domestic demand increases. In addition, inadequate supply of plasma protein is a frequent problem. Also, due to concern of contamination of host animals through the feeding of plasma protein there is becoming a further need for a non-plasma product which can be utilized in those formulations where plasma protein is either not available or its use barred.
In addition, it is well known that protein hydrolyzates prepared by reaction of selected protein materials with at least one proteolytic enzyme can also be used in feed formulations. Such compounds are generally in such amounts as to maximize its content in the feed formulation without having a negative effect on the host. Such hydrolyzates provided a replacement for fish meal as a protein source and thus while providing nutritional values provided no other benefit in the growth of the host.
Thus there is presently experienced a need for a suitable replacement for plasma as an ingredient of feed formulations as well as the need to achieve growth enhancement for host animals.
The aforementioned need is met by the present invention by providing a bioactive peptide composition which not only provides an alternative to plasma products but also serves to enhance the growth of both animals and fish.
According to the present invention a bioactive peptide composition is produced by the enzymatic hydrolysis of a protein using the proteolytic enzyme derived from cod, specifically the stomachs of Atlantic cod.
The present invention is based upon the discovery that the peptide product obtained through enzymatic hydrolysis of a protein source with the pepsin enzyme derived from the stomach of Atlantic cod can be used as a substitute for plasma protein in achieving growth enhancement of warm blooded animals.
In a first aspect of the present invention there is provided novel animal feed compositions which enhance growth.
In addition, the present invention is also based on the further discovery that such peptide products can be used at low levels to enhance the growth of fish.
Accordingly, in one aspect of the present invention there is provided a novel peptide product which is useful for enhancing the growth of warm blooded animals and fish.
In another aspect of this invention there is provided a process for the production of growth enhancing peptides from a protein source.
In another aspect of this invention there is provided a novel process for enhancing the growth of warm blooded animals and fish.
Other aspects, objects and the several advantages of this invention will be apparent from the foregoing disclosure and appended claims.
The term bioactive peptide compositions as used herein is intended to define a composition consisting essentially of a mixture of peptides with aromatic amino acids in N-terminal position, produced by hydrolysis of a protein source at a pH of 2-6 with pepsins from fish as the hydrolytic enzyme.
The bioactive peptide product of this invention is prepared by a process comprising the steps of:
(a) hydrolyzing a selected protein material, dispersed in an aqueous medium, together with cod pepsin at a controlled temperature and acidic pH;
(b) removing lipids and solids from the pepsin treated acidized protein source; and thereafter
(c) recovering the resulting bioactive peptide.
The process may employ protein material from any of a great variety of sources including meats, fish and plants. Protein material from fish is particularly preferred for use in the process of this invention.
The process of this invention is particularly adapted to effecting protein hydrolysis in an acidic medium of controlled pH to produce a bioactive peptide composition suitable for use in the enhancement of growth of host animals. Protein material is prepared for treatment by grinding, comminution, or other means to provide a suitable particle size for slurring with acid. The aqueous slurry is heated with agitation at a combination of temperature and time such that the initial reaction velocity of hydrolysis is maximized.
The enzyme employed in the process of this invention is the proteolytic pepsin enzyme derived from the stomach of Atlantic cod. In one presently preferred embodiment of this invention the enzyme is obtained by a process which comprises homogenizing the cod stomach in the presence of enough formic acid to lower the pH to 4 and then heating for a period of time whereby the homogenate becomes a liquid slurry due to the digestion of the tissues by the enzymes present in the stomach and thereafter recovering from the aqueous phase following removal of sludge and oils following concentration and drying the desired pepsin enzyme.
In carrying out the process, the homogenate of the Atlantic cod stomach can also be employed as the enzyme source. Thus following the homogenizing of the cod stomach in the presence of acid and the digestion of the tissues thereof by the enzymes present therein, the resulting liquid slurry which is enzyme rich can be employed directly into the process of the present invention as the enzyme source for the production of bioactive peptides.
In the production of the bioactive peptide composition of the invention, a suitable protein source is treated at a pH in the range of about 2-6, preferably at a pH of 4, with the pepsin enzyme derived from the Atlantic cod stomachs for a time to effect hydrolysis of the protein source so as to form a mixture of peptides having aromatic amino acids in N-terminal positions which following heating and thereafter removal of oils and sludge provided a liquid composition consisting essentially of bioactive peptides and deactivated cod pepsin.
The hydrolysis step is conducted in a continuous stirred-tank reactor employing an enzyme to protein substrate ratio selected to obtain the desired level of conversion within a time period of 24 to 100 hours.
Generally the enzyme is added in amounts in the range of 0.5 to 5 g per 1000 kg of protein.
Acid employed in the hydrolysis process in an amount to provide normally 3% volume by volume.
Following the removal of oils and undissolved solids from the partially hydrolyzed aqueous protein material dispersion, the recovered protein hydrolyzate fraction may be concentrated, as by reverse osmosis or thermal evaporation and dried, as by freeze drying or spray drying, and sent to product storage for subsequent packaging and distribution.
Pepsin enzyme suitable for use in one embodiment of the present invention is obtained by hydrolysis of Atlantic cod fish stomachs at a pH in the range of 2-6. Following hydrolysis, the resulting hydrolyzate is subjected to lipid and solids removal followed by ultrafiltration of the resulting liquid peptide pepsin mixture to effect recovery of the pepsin enzyme therefrom.
The Atlantic cod pepsin which is used in the process of this invention is a proteolytic enzyme prepared from Atlantic cod (Gadus morhua) stomach by autolysis. The enzyme has a specific activity of 100-200 Anson Units expressed as xcexc mol TCA-soluble tyrosine released from haemoglobin per hour at pH 3 and 25xc2x0 C.
The compositions produced by the process of this invention have been found to possess significantly distinctive properties. Such compositions are bioactive peptides which are capable of enhancing growth when included in feed compositions for host animals.
Accordingly, the bioactive peptides of this invention are most appropriate for providing a process for the enhancement of growth of an animal by feeding such animal with an amount of such bioactive peptide to effect growth enhancement of the host animal. In general such growth enhancement is achieved when the bioactive peptide is introduced into the host animal in an amount in the range of 0.1 to 5 g/kg body weight per day.
Feed formulations using the active peptide compositions of this invention will be added to conventional feed ingredients for the intended host animal an amount of bioactive peptide which is sufficient to achieve growth enhancement activity.
Generally such amounts will be in the range of from about 0.1 to about 5 weight percent of the total feed, preferably from about 1 to 2%.
Enzyme hydrolysis of the protein source is normally carried out at a temperature in the range of about 10xc2x0 C. to about 45xc2x0 C. and at a pH in the range of about 2 to about 6. Suitable pH is achieved by use of either a mineral or organic acid or salts thereof which is added prior to addition of the pepsin. The amount of acid employed is that which will provide the desired pH level.
In one presently preferred embodiment of the invention there is employed a lower carboxylic acid namely formic acid.
Protein source starting materials suitable for the process of the present invention are:
1. fish wastes including whole trash fish, fish left after filleting, fish solubles, fish viscera and any other materials which are by-products of the fishing industry and processing
2. pork skins and tissues
3. beef tissue
4. soy bean proteins or other seed proteins
5. milk proteins
The following examples will further illustrate the process and products of the invention.